Design of Vibrio 16S rRNA Gene Specific Primers and Their Application in the Analysis of Seawater Vibrio Community

类 Vibrio 原因 vibriosis 的病原的种， maricultured 动物和海味消费者的最流行的疾病之一。在海味生产的链监视他们的动力学，处理并且消费为食物和海洋生物养殖安全是很重要的。为了充实代表 Vibrio 16S ribosomal RNA 基因(rDNA ) ，碎裂并且识别进一步真实及时的这些细菌并且同时地在在链的细菌的植物群之中，有选择地放大 Vibrio 16S rDNA 碎片的一双教材与他们在海水 Vibrio 的分析证明的特性和报道被设计社区。二份教材， VF169 和 VR744 的 Thespecificities 和范围，被使用强风节目在在 GenBank 可得到的细菌的 16S rDNAs 之中理论上决定，实际上，由放大 Vibrio16S， rDNA 从海水 DNA 碎裂。超过 88.3% 序列在 GenBank，与 VR744 显示出相同火柴，属于 Vibrio 类。33 克隆的一个总数随机被选择并且定序。所有序列显示出他们的最高的类似到并且多样的已知的 Vibrio 种类在那些附近聚类。设计的教材能够检索宽范围 ofVibrio 16S rDNA 在海水在细菌的植物群之中明确地碎裂，海味耕作的最重要的自然环境。

Genetic Diversity of Three Aristichthys nobilis Populations and One Inbreeding Stock

Genetic diversity of threeAristichthys nobilis populations and one inbreeding stock was studied using random amplified polymorphic DNA (RAPD) method. Materials of natural populations were collected from two counties of Guangxi Province, Hengxian (NH) and Shanglin (NS) and one city of Hubei Province, Wuhan city. (NW). Shanglin and Hengxian's river system respectively belongs to Qianjiang and Yujiang River, two tributaries confluencing to Xunjiang River, main tributary of Xijiang River, which is the biggest tributary of the Zhujiang River system, and Wuhan's water system belongs to the Yangtse River system. The inbreeding stock (RS) was the more than 10 generation descendant by brother-sister mating system whose parents were collected from Shanglin. The results showed that the genetic variety of individuals of RS was very low (0.105 4), whereas that of natural populations was relatively high, which from high to low was 0.158 1 (NW), 0.132 0 (NH) and 0.110 5 (NS). As an index for genetic distance pair-wise populations, the genetic variety between populations was studied, which characterized the genetic distance between populations. The genetic distance of NW and NH, NW and NS, NW and RS were respectively high, whereas that of NH and NS, NS and RS, NH and RS was low. Chi-test (χ 2) and the analysis of the genetic variety pair-wise populations was taken as efficient approach for studying population difference.

Identification of Prorocentrum minimum and Takayama pulchella by fluorescence in situ hybridization through epifluorescence microscopy and flow cytometry

Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified, cloned and sequenced, and these sequence data were deposited in the GenBank. Eight oligonucleotide probes （DNA probes） were designed based on the sequence analysis. The probes were employed to detect and identify P. minimum and T. pulchella in unialgal and mixed algal samples with a fluorescence in situ hybridization method using flow cytometry. Epifluorescence micrographs showed that these specific probes labeled with fluorescein isothiocyanate entered the algal cells and bound to target sequences, and the fluorescence signal resulting from whole-cell hybridization varied from probe to probe. These DNA probes and the hybridization protocol we developed were specific and effective for P. minimum and T. pulchella, without any specific binding to other algal species. The hybridization efficiency of different probes specific to P. minimum was in the order： PM18S02 PM28S02 〉 PM28S01 〉PM18S01, and that of the probes specific to T. pulcheUa was TP18S02 TP28S01 〉 TP28S02 〉TP18S01. The different hybridization efficiency of the DNA probes could also be shown in the fluorescent signals between the labeled and unlabeled cells demonstrated using flow cytometry. The DNA probes PM18S02, PM28S02, TP18S02 and TP28S01, and the protocol, were also useful for the detection of Mgae in natural samples.